Isobaric Mass Differential Labeling Reagents for Multiplex Analysis
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Solution Overview
Problem
Current methods for determining analytes by mass spectrometry face challenges in accurately quantifying and identifying multiple analytes in complex mixtures, particularly in multiplex analyses, due to limitations in distinguishing between isobaric and mass-differential compounds.
Innovation Solution
The use of isobaric and mass-differential labeling reagents, which are sets of compounds that react with analytes to form stable adducts, allowing for unique identification and quantification through mass spectrometry by generating distinct reporter ions, enabling simultaneous analysis of multiple samples and compounds with the same or different masses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If isobaric labeling reagents are used to label multiple analytes, then the quantification capability is improved, but the ability to distinguish between different labeled analytes deteriorates because they have the same mass
Solution Approach 1:
The labeling reagent is divided into three distinct segments: a reporter group that provides a unique mass signature, a balance group that adjusts the overall mass, and a reactive group that attaches to the analyte. This segmentation allows isobaric reagents to be distinguished by their reporter group masses while maintaining the same total reagent mass, resolving the contradiction between quantification capability and distinguishability.
Solution Approach 2:
Different portions of the isobaric labeling reagents have different local properties: the reporter group has a unique mass that serves as an identifier, while the balance group compensates to maintain isobarity. This local differentiation allows the reagents to function both as quantification labels and as distinguishable entities in mass spectrometry analysis.
2Difficulty of detecting and measuring
If mass differential labeling reagents are used, then the distinction between different analytes is improved, but the quantification accuracy deteriorates due to mass variations affecting detection
Solution Approach 1:
The invention controls the mass parameter of the balance group to compensate for variations in the reporter group mass. By adjusting the balance group's mass, the total reagent mass is maintained at a constant value, ensuring that mass differential reagents produce consistent detection signals while still allowing distinction through their unique reporter groups.
3Adaptability or versatility
If multiple labeling reagents are used for multiplex analysis, then the analysis capability is improved, but the system complexity deteriorates due to managing multiple reagents and their combinations
Solution Approach 1:
The labeling reagent structure is designed to be universal across multiple analyte types. The standardized three-component structure (reporter, balance, reactive group) can be applied to label different analytes in a multiplexed manner, allowing a single systematic approach to handle multiple samples and analyte classes without requiring fundamentally different reagent designs.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables precise relative and absolute quantification of analytes in complex samples, facilitating multiplex analysis and improving the accuracy of protein and small molecule identification, such as in proteomic and genomic studies, by utilizing unique reporter ions to distinguish between differently labeled analytes.
Implementation Method 1
react with an amine or hydrazine group to thereby form a stable adduct
Data Source
AI summary
This invention pertains to methods, mixtures, kits and compositions pertaining to analyte determination by mass spectrometry using labeling reagents that comprise a nucleophilic reactive group that reacts with a functional group of an analyte to produce a labeled analyte. The labeling reagents can be used as isobaric sets, mass differential labeling sets or in a combination of isobaric and mass differential labeling sets.